The fish were starved for 24 hours before blood samples were taken. Three drops of the commercial clove oil extract dissolved in 10 L of tap water were used to anaesthetize fish (n = 6 per treatment). Whole blood samples were taken from the caudal peduncle and then placed in tiny plastic vials containing heparin to determine the hematological parameters. Additional blood samples were taken to obtain the serum by centrifuging the blood without heparin at 3500 g for 20 minutes, then stored in a deep freezer (-20°C) until biochemical analysis.
(1) Hematological Parameters. Red blood cells (RBCs), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), platelets (PLT), packed cell volume (PCV), and total white blood cells (WBCs) were all measured in the whole blood samples. The measurement of Hb (mg dL−1) was done using commercial colorimetric kits (Diamond Diagnostic, Egypt). Using the methods of Dacie and Lewis [36 ], PLT (>103 mm−3) and RBCs (>106 mm−3) were counted using an Ao Bright-Line hemocytometer model (Neubauer Enhanced, Precicolor HBG, Germany). MCV and MCHC (%) were computed using the prescribed methods of Beutler et al. [37 ] while PCV (%) was measured based on the methods of Stoskopf [38 ].
(2) Serum Biochemical Traits. Utilizing commercial kits (Diagnostic System Laboratories, Inc., USA), serum biochemical components such as alanine transaminase (ALT), aspartate transaminase (AST), uric acid (UA), creatinine, total protein (TP), albumin (ALB), and globulin (GLB) were calorimetrically evaluated. TP (g dL−1) and ALB (g dL−1) were measured according to McGowan et al. [39 (link)] whereas serum ALT (U L−1) and AST (U L−1) were assessed in accordance to the methods of Henry [40 ]. Serum GLB (g dL−1) were calculated using the variations between TP and ALB. Triiodothyronine (T3, ng dL−1) and thyroxine (T4, g dL−1) serum concentrations were also measured using the Cobas 6000 immunoassay analyzer test and commercial RIA kits (Roche, Basel, Switzerland).
(1) Hematological Parameters. Red blood cells (RBCs), hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC), platelets (PLT), packed cell volume (PCV), and total white blood cells (WBCs) were all measured in the whole blood samples. The measurement of Hb (mg dL−1) was done using commercial colorimetric kits (Diamond Diagnostic, Egypt). Using the methods of Dacie and Lewis [36 ], PLT (>103 mm−3) and RBCs (>106 mm−3) were counted using an Ao Bright-Line hemocytometer model (Neubauer Enhanced, Precicolor HBG, Germany). MCV and MCHC (%) were computed using the prescribed methods of Beutler et al. [37 ] while PCV (%) was measured based on the methods of Stoskopf [38 ].
(2) Serum Biochemical Traits. Utilizing commercial kits (Diagnostic System Laboratories, Inc., USA), serum biochemical components such as alanine transaminase (ALT), aspartate transaminase (AST), uric acid (UA), creatinine, total protein (TP), albumin (ALB), and globulin (GLB) were calorimetrically evaluated. TP (g dL−1) and ALB (g dL−1) were measured according to McGowan et al. [39 (link)] whereas serum ALT (U L−1) and AST (U L−1) were assessed in accordance to the methods of Henry [40 ]. Serum GLB (g dL−1) were calculated using the variations between TP and ALB. Triiodothyronine (T3, ng dL−1) and thyroxine (T4, g dL−1) serum concentrations were also measured using the Cobas 6000 immunoassay analyzer test and commercial RIA kits (Roche, Basel, Switzerland).
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